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Targeted Transcriptional Profiling of Microdissected Biopsy Specimens Representing Early Colonic Neoplasia
Author(s) -
Mo Allen,
Jackson Stephen,
Devers Thomas J.,
Rosenberg Daniel W.
Publication year - 2016
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25644
Subject(s) - laser capture microdissection , microdissection , stromal cell , biology , rna , biopsy , computational biology , pathology , cancer research , gene expression , medicine , gene , genetics
Our incomplete understanding of the critical changes that accompany the earliest stages of tumor initiation provides a substantial hurdle for the development of novel intervention strategies for cancer prevention. Premalignant lesions are inherently difficult to characterize given their diminutive size, creating technical obstacles for accurate genetic profiling. Here, we describe an approach combining laser‐capture microdissection (LCM) with targeted RNA‐sequencing to study the transcriptional state of epithelial and stromal cells during the earliest detectable stage of human colorectal neoplasia, the aberrant crypt foci (ACF). We provide a robust and reproducible workflow for RNA isolation, library preparation, and expression profiling of laser‐captured cells from frozen OCT‐embedded tissue specimens. It is anticipated that the methodological approach outlined in this report will provide a framework for a broad range of microgenomics analyses that can be routinely applied to many other premalignant tissues. J. Cell. Biochem. 117: 2677–2681, 2016. © 2016 Wiley Periodicals, Inc.